Project Summary/Abstract New technology is needed to investigate nanoscale intercellular communication in dendritic cells, a process which shares commonality with a broad range of tissues such as the skeletal, nervous, cardiovascular, and immune systems. The proposed work will design, build and characterize an integrated multiscale fabrication and imaging platform capable of building multiscale devices and systems using biocompatible and commonly used materials in biomedical sciences, and capable of non-invasively imaging the biology-of-interest at multiple scales with high resolution and high speed. This project will be divided into two specific aims, with two modules in each aim. Aim 1 will focus on developing the capability of fabricating devices that could span from centimeter to nanometer resolution using commonly used polydimethylsiloxane, synthetic polyethylene glycol diacrylate hydrogel and naturally derived gelatin methacrylate hydrogel materials. In module 1, a hybrid additive-subtractive process based on ultrafast lasers will be developed to allow fabrication of devices with a feature range of several centimeters to 0.5micrometer, while in module 2, STimulated Emission Depletion (STED) inspired lithography will be used to print nanoscale structures with a feature range from 0.5micrometer to less than 100nanometers. Aim 2 will focus on integrating super resolution and 3D sectional imaging capabilities within the platform. In module 3, STED microscopy will be designed to achieve an imaging resolution of sub-100nm, while in module 4, Digital Micromirror Device-Structural Illumination Microscopy (DMD-SIM) will be used to achieve high-speed wide-angle sectional imaging capabilities. We will demonstrate the new capabilities of the proposed platform by developing a multiscale fluidic device that replicates the 3D nanoscale canaliculi structure found in the skeletal systems ? a task not possible with current technology. If successful, the proposed work will enable researchers to ask new questions concerning a broad range of cells, tissues, systems and disease types that could not be studied adequately in the absence of such a technology.